WO2025181244A1

WO2025181244A1 - Use of probiotic combinations for alleviating menopausal symptoms

Info

Publication number: WO2025181244A1
Application number: PCT/EP2025/055357
Authority: WO
Inventors: Maija Emilia MARTTINEN; Pia Johanna M. MAUKONEN; Neeta SENGUPTA; Liisa LEHTORANTA; Reeta ALA-JAAKKOLA
Original assignee: International N&H Denmark ApS
Current assignee: International N&H Denmark ApS
Priority date: 2024-02-28
Filing date: 2025-02-27
Publication date: 2025-09-04
Anticipated expiration: 2026-08-28

Abstract

The present invention relates to therapeutic and non-therapeutic use of combinations of bacterial strains. In particular the use is for preventing or alleviating symptoms of menopause or climacteric symptom in a female mammal. Also disclosed herein are specific combinations of bacterial strains and their use in, as well as methods for reducing the need for estrogen replacement therapy, or for use in combination with estrogen replacement therapy, or other menopausal hormone replacement therapies.

Description

USE OF PROBIOTIC COMBINATIONS FOR ALLEVIATING MENOPAUSAL SYMPTOMS

FIELD OF THE INVENTION

BACKGROUND OF THE INVENTION

Menopause is clinically defined as the end of the reproductive period and diagnosed after 12 months without a menstrual period; however, menopausal symptoms are experienced during both the menopausal transition (perimenopause) and post-menopause stage. Levels of estrogens and progesterone begin to fluctuate already before menopause during the menopausal transition, also referred to as perimenopause, which lasts on average 4.5 years (Woods et al. 2002). Menopausal symptoms, such as hot flushes, nights sweats, insomnia, changes in mood, vaginal dryness, and reduced libido, occur in perimenopause, menopause and post-menopause, with perimenopause being responsible for the majority of symptoms. Menopausal symptoms are unique to all women. They can vary from mild to severe, be non-existent or persist for years. Vasomotor symptoms (VMS) can be considered a hallmark of menopause and they are experienced by approximately 75% of women during perimenopause and at post-menopause (Monteleone et al. 2018). Vasomotor symptoms are also reported as the most troublesome and frequent symptoms that can influence quality of life during the daytime but also at night, and thus affect the sleep quality. Sleep disorders are important complaints in postmenopause and in menopausal transition (Monterrosa-Castro et al. 2016). Menopausal symptoms and VMS are associated with increased stress levels (Weidner et a/. 2020). Anxiety symptoms are increased in menopause and during menopausal transition (Monteleone et al. 2018) and may be associated with symptom severity. Somatic anxiety has been reported to be associated with VMS (Freeman and Sammel 2016). Menopausal hot flush is the major menopausal symptom that manifests as an increase in skin temperature (Sturdee et al. 2017). Decline is systemic estrogen level and other hormonal changes during the menopausal transition, lead to narrowing of the thermoneutral zone in the central nervous system: a minor change in the core body temperature triggers a neuroendocrine response to maintain the core temperature and vasomotor flushing is the outcome of this compensatory mechanism (Zhang et al. 2021). Nocturnal hot flushes are called night sweats. Ovariectomized (OVX) rat model is well characterized in the scientific community, and it is the most researched model to study menopause related symptoms and conditions associated with estrogen deficiency/decline in humans (Koebele and Bimonte-Nelson 2016; Medina-Contreras et al. 2020; Yousefzadeh et al. 2020). Ovariectomy results in depletion of systemic estrogens and ovariectomy-induced changes reported in OVX rodents mimic the physiological changes associated with human menopause (i.e. body weight gain, increased risk for osteoporosis and metabolic syndrome, regulation of vasomotor symptoms). Measuring tail skin temperature in OVX rat is well-documented and accepted model for menopausal hot flushes when screening pharmacological compounds and other ingredients for symptom alleviation (Kobayashi et al. 2000; Opas et al. 2004; Williams et al. 2010). The increase in the tail temperature due to estrogen deficiency in OVX rats is similar to hot flushes in humans. In women, menopausal hormone replacement therapies provide relieve to hot flushes and similarly administration of 170-estradiol (E2) normalized the OVX induced increase in tail temperature (Kobayashi et al. 2000).

EP1186295 relates to pharmaceutical preparations containing extracts of soy isoflavone and probiotic micro-organisms.

US20190192591 relates to a Lactobacillus acidophilus YT1 strain for treating menopause. W0202091166 relates to Lactobacillus plantarum NK3 and Bifidobacterium longum NK49 for the treatment of menopausal disease in women.

WO2022130317 disclose the use of probiotic compositions comprising a strain belonging to the Lactobacillus reuteri species in association with a strain belonging to the species of Bifidobacterium breve or Bifidobacterium lactis, for use in the treatment of pathological conditions related to stress.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1. Total body weight at sacrifice (g). Statistical analysis was performed using Kruskal-Wallis test. As statistical differences were observed (p=< 0.001), the pairwise comparisons were performed using Dunn's test. Notation: 'COMP'=Comparison group, '***'=p-value < 0.001, '*' = p- value < 0.05, 'NS' = non-significant.

Figure 2. Uterus weight, wet (mg). Prior to statistical analysis the data were transformed using logarithmic transform. Statistical analysis was performed using ANOVA. As statistical differences were observed (p=< 0.001), the pairwise comparisons were performed using ANOVA contrasts.

Notation: 'COMP'=Comparison group, '***'=p-value < 0.001, '*' = p-value < 0.05, 'NS' = non-sig- nificant. Figure 3. Tail skin temperature (Ttaii) at day 84 (°C). Statistical analysis was performed using Kruskal-Wallis test. As statistical differences were observed (p=< 0.001), the pairwise comparisons were performed using Dunn's test. Notation: 'COMP'=Comparison group, '***'=p-value < 0.001, '**'=p-value < 0.01, '*'=p-value < 0.05.

SUMMARY OF THE INVENTION

It is an object of embodiments of the invention to provide improved compositions and methods for treating, preventing or alleviating the symptoms of one or more symptom of menopause or climacteric symptom in a female mammal, such as a female human.

The present invention relates in a broad aspect to a composition of bacterial strains for use in treating, preventing and/or alleviating one or more symptom of menopause or climacteric symptom in a female mammal, such as a female human.

Accordingly, in a first aspect the present invention relates to a composition comprising a combination of at least one bacterial strain of Bifidobacterium animaiis subsp. lactis in combination with at least one bacterial strain selected from a strain of Lacticaseibacillus paracasei and/or a strain of Lactiplantibacillus plantarum for use in treating, preventing and/or alleviating one or more symptom of menopause or climacteric symptom in a female mammal, such as a female human.

In a second aspect the present invention relates to a composition of a bacterial strain of Bifidobacterium animaiis subsp. lactis in combination with a bacterial strain of Lacticaseibacillus paracasei and/or Lactiplantibacillus plantarum for use in reducing the need for estrogen replacement therapy, or for use in combination with estrogen replacement therapy, or other menopausal hormone replacement therapies.

In a third aspect the present invention relates to the use of a composition of a bacterial strain of Bifidobacterium animaiis subsp. lactis in combination with a bacterial strain of Lacticaseibacillus paracasei and/or Lactiplantibacillus plantarum for the manufacture of a food product, a dietary supplement or a pharmaceutically acceptable formulation for use in treating, preventing and/or alleviating one or more symptom of menopause in a female mammal, such as a female human.

In a further aspect the present invention relates to a method for treating, preventing and/or alleviating one or more symptom of menopause in a female mammal, such as a female human comprising the steps of: a) providing a composition of at least one bacterial strain of Bifidobacterium animaiis subsp. lactis in combination with at least one bacterial strain selected from a strain of Lacticaseibacillus paracasei and/or a strain of Lactiplantibacillus plantarum,- and b) administering said composition of strains to said female mammal under conditions such that the risk of developing one or more symptom of menopause is reduced. In some embodiments the composition of at least one bacterial strain of Bifidobacterium animaiis subsp. lactis in combination with at least one bacterial strain selected from a strain of Lacticaseibacillus paracasei and/or a strain of Lactiplantibacillus plantarum for use in the method according to the present invention is as defined herein.

DETAILED DESCRIPTION OF THE INVENTION

The present inventors have found that specific combinations of bacterial strains have a significant and improved effect in treating, preventing and/or alleviating one or more symptom of menopause or climacteric symptom in a female mammal, such as a female human.

The present patent examples disclosed herein demonstrated a reduction in tail skin temperature, a proxy measure for hot flushes, with supplementation of probiotic compositions of a strain of Bifidobacterium animaiis subsp. lactis in combination with at least one bacterial strain selected from a strain of Lacticaseibacillus paracasei and/or a strain of Lactiplantibacillus plantarum. In particular a reduction in tail skin temperature was seen with the specific combinations of Bifidobacterium animaiis subsp. lactis B420 + Lacticaseibacillus paracasei Lpc-37; B. animaiis subsp. lactis B420 + Lactiplantibacillus plantarum Lp-115; and B. animaiis subsp. lactis BI-04 + L. plantarum Lp-115) in the OVX rat model.

Bacteria

The bacterial strains used in aspects of the invention are bacterial strains of the species Bifidobacterium animaiis subsp. lactis, Lacticaseibacillus paracasei, and Lactiplantibacillus plantarum. In one aspect, the Bifidobacterium animaiis subsp. lactis is strain B420 or BI-04; in another aspect, Lacticaseibacillus paracasei is strain Lpc-37; in another aspect, Lactiplantibacillus plantarum is strain Lp-115; These strains are commercially available from DuPont Nutrition Biosciences ApS, previously Danisco A/S, now International Flavors & Fragrances Inc.

Bifidobacterium animaiis subsp. lactis B420, is deposited at DSMZ on 30 June 2015 under registration number DSM 32073. Lactiplantibacillus plantarum Lp-115 deposited at DSMZ on 9 February 2009 under registration number DSM 22266. Bifidobacterium lactis BI-04 deposited on 19 May 2020 at DSMZ under registration number DSM 33525. Lacticaseibacillus paracasei Lpc-37 deposited on 5 October 2017 at DSMZ under registration number DSM 32661.

Preferably the bacterial strains used in the present invention are bacterial strains which are generally recognised as safe (GRAS) and, which are preferably GRAS approved. GRAS is an American Food and Drug Administration (FDA) designation that a chemical or substance added to food is considered safe by experts, and so is exempted from the usual Federal Food, Drug, and Cosmetic Act (FFDCA) food additive tolerance requirements. The bacterial strains, when used in aspects of the invention, are suitable for human and/or animal consumption. A skilled person will be readily aware of specific strains which are used in the food and/or agricultural industries and which are generally considered suitable for human and/or animal consumption.

Optionally, the bacterial strains when used in aspects of the invention are probiotic bacteria. The term "probiotic bacteria" is defined as covering any non-pathogenic bacteria which, when administered live in adequate amounts to a host, confers a health benefit on that host. For classification as a "probiotic", the bacteria must survive passage through the upper part of the digestive tract of the host. They are non-pathogenic, non-toxic and exercise their beneficial effect on health on the one hand via ecological interactions with the resident flora in the digestive tract, and on the other hand via their ability to influence the host physiology and immune system in a positive manner. Probiotic bacteria, when administered to a host in sufficient numbers, have the ability to progress through the intestine, maintaining viability, exerting their primary effects in the lumen and/or the wall of the host's gastrointestinal tract. They then transiently form part of the resident flora and this colonisation (or transient colonisation) allows the probiotic bacteria to exercise a beneficial effect, such as the repression of potentially pathogenic micro-organisms present in the flora and interactions with the host in the intestine including the immune system.

Thus, in a particular aspect of the present invention, the bacterial strains for use according to the invention is a probiotic strain.

Compositions

The term "composition" is used in the broad sense to mean the way something is composed, i.e. its general makeup. In aspects of the invention, the compositions may consist essentially of a single strain of the species Lacticaseibacillus paracasei bacteria.

Alternatively, the compositions may comprise bacterial strains according to the present invention together with other components, such as biological and chemical components, active ingredients, metabolites, nutrients, fibres, prebiotics, etc.

In one aspect, the present invention provides for the use of composition comprising a combination of at least one bacterial strain of Bifidobacterium animaiis subsp. lactis in combination with at least one bacterial strain selected from a strain of Lacticaseibacillus paracasei and/or a strain of Lactiplantibacillus plantarum, wherein said bacterial strains are administered in the form of compositions, such as food products, food ingredients, functional foods, dietary supplements, and pharmaceutically acceptable formulations. In a particular aspect, the compositions according to the present invention further comprise prebiotics.

In yet a further aspect of the present invention, the bacterial strains according to the present invention are present in the composition in an amount between 10⁶ and 10¹², e.g. between 10⁸ and 10¹² colony forming units (CFU) per dose, optionally 10¹⁰ CFU per dose. While it is not a requirement that the compositions comprise any support, diluent or excipient, such a support, diluent or excipient may be added and used in a manner which is familiar to those skilled in the art. Examples of suitable excipients include, but are not limited to, microcrystalline cellulose, rice maltodextrin, silicone dioxide, and magnesium stearate. The compositions of the invention may also comprise cryoprotectant components (for example, glucose, sucrose, lactose, trehalose, sodium ascorbate and/or other suitable cryoprotectants).

The terms "composition" and "formulation" may be used interchangeably.

Compositions used in aspects of the invention may take the form of solid, liquid, solution or suspension preparations. Examples of solid preparations include, but are not limited to: tablets, pills, capsules, granules and powders which may be wettable, spray-dried or freeze dried/ly- ophilized. The compositions may contain flavouring or colouring agents. The compositions may be formulated for immediate-, delayed-, modified-, sustained-, pulsed- or controlled-release applications.

By way of example, if the compositions of the present invention are used in a tablet form, the tablets may also contain one or more of: excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate and glycine; disintegrants such as starch (preferably corn, potato or tapioca starch), sodium starch glycollate, croscarmel- lose sodium and certain complex silicates; granulation binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin and acacia; lubricating agents such as magnesium stearate, stearic acid, glyceryl behenate and talc may be included.

Examples of other acceptable carriers for use in preparing compositions include, for example, water, salt solutions, alcohol, silicone, waxes, petroleum jelly, vegetable oils, polyethylene glycols, propylene glycol, liposomes, sugars, gelatin, lactose, amylose, magnesium stearate, talc, surfactants, silicic acid, viscous paraffin, perfume oil, fatty acid monoglycerides and diglycerides, hydroxymethylceilulose, polyvinylpyrrolidone, and the like.

For aqueous suspensions and/or elixirs, the composition of the present invention may be combined with various sweetening or flavouring agents, colouring matter or dyes, with emulsifying and/or suspending agents and with diluents such as water, propylene glycol and glycerin, and combinations thereof.

Specific non-limiting examples of compositions which can be used in aspects of the invention are set out below for illustrative purposes. These include, but are not limited to food products, food ingredients, functional foods, dietary supplements, pharmaceutical compositions and medicaments. Accordingly, in some embodiments, the compositions of the present invention are used for non-therapy or in a non-therapeutic composition. In other embodiments, the compositions of the present invention are used for therapy or in a medical composition or medicament. Food products

The compositions of the invention may take the form of a food product. Here, the term "food" is used in a broad sense and covers food and drink for humans as well as food and drink for animals (/'.e. a feed). Preferably, the food product is suitable for, and designed for, human consumption.

The food may be in the form of a liquid, solid or suspension, depending on the use and/or the mode of application and/or the mode of administration.

When in the form of a food product, the composition may comprise or be used in conjunction with one or more of: a nutritionally acceptable carrier, a nutritionally acceptable diluent, a nutritionally acceptable excipient, a nutritionally acceptable adjuvant, a nutritionally active ingredient.

By way of example, the compositions of the invention may take the form of one of the following : A fruit juice; a beverage comprising whey protein: a health or herbal tea, a cocoa drink, a milk drink, a lactic acid bacteria drink, a yoghurt and/or a drinking yoghurt, a cheese, an ice cream, a water ice, a dessert, a confectionery, a biscuit, a cake, cake mix or cake filling, a snack food, a fruit filling, a cake or doughnut icing, an instant bakery filling cream, a filling for cookies, a ready-to-use bakery filling, a reduced calorie filling, an adult nutritional beverage, an acidified soy/juice beverage, a nutritional or health bar, a beverage powder, a calcium fortified soy milk, or a calcium fortified coffee beverage.

Optionally, where the product is a food product, the bacterial strains of Bifidobacterium animalis subsp. lactis in combination with at least one bacterial strain selected from a strain of Lacticaseibacillus paracasei and/or a strain of Lactiplantibacillus plantarum should remain effective through the normal "sell-by" or "expiration" date during which the food product is offered for sale by the retailer. Preferably, the effective time should extend past such dates until the end of the normal freshness period when food spoilage becomes apparent. The desired lengths of time and normal shelf life will vary from foodstuff to foodstuff and those of ordinary skill in the art will recognize that shelf-life times will vary upon the type of foodstuff, the size of the foodstuff, storage temperatures, processing conditions, packaging material and packaging equipment.

Food ingredients

Compositions of the present invention may take the form of a food ingredient and/or feed ingredient.

As used herein the term "food ingredient" or "feed ingredient" includes a composition which is or can be added to functional foods or foodstuffs as a nutritional and/or health supplement for humans and animals.

The food ingredient may be in the form of a liquid, suspension or solid, depending on the use and/or the mode of application and/or the mode of administration. Functional Foods

Compositions of the invention may take the form of functional foods.

As used herein, the term "functional food" means food which is capable of providing not only a nutritional effect but is also capable of delivering a further beneficial effect to the consumer.

Accordingly, functional foods are ordinary foods that have components or ingredients (such as those described herein) incorporated into them that impart to the food a specific function - e.g. medical or physiological benefit - other than a purely nutritional effect.

Although there is no legal definition of a functional food, most of the parties with an interest in this area agree that they are foods marketed as having specific health effects beyond basic nutritional effects.

Some functional foods are nutraceuticals. Here, the term "nutraceutical" means a food which is capable of providing not only a nutritional effect and/or a taste satisfaction but is also capable of delivering a therapeutic (or other beneficial) effect to the consumer. Nutraceuticals cross the traditional dividing lines between foods and medicine.

Dietary Supplements

The compositions of the invention may take the form of dietary supplements or may themselves be used in combination with dietary supplements, also referred to herein as food supplements.

The term "dietary supplement" as used herein refers to a product intended for ingestion that contains a "dietary ingredient" intended to add nutritional value or health benefits to (supplement) the diet. A "dietary ingredient" may include (but is not limited to) one, or any combination, of the following substances: bacteria, a probiotic (e.g. probiotic bacteria), a vitamin, a mineral, a herb or other botanical, an amino acid, a dietary substance for use by people to supplement the diet by increasing the total dietary intake, a concentrate, metabolite, constituent, or extract.

Dietary supplements may be found in many forms such as tablets, capsules, soft gels, gel caps, liquids, or powders. Some dietary supplements can help ensure an adequate dietary intake of essential nutrients; others may help reduce risk of disease.

Pharmaceutical compositions (formulations)

Compositions of the invention may be used as - or in the preparation of -pharmaceuticals. Here, the term "pharmaceutical" is used in a broad sense - and covers pharmaceuticals for humans as well as pharmaceuticals for animals (/.e. veterinary applications).

In a preferred aspect, the pharmaceutical is for human use. The pharmaceutical can be for therapeutic purposes - which may be curative, palliative or preventative in nature.

A pharmaceutical may be in the form of a compressed tablet, tablet, capsule, ointment, suppository or drinkable solution.

When used as - or in the preparation of - a pharmaceutical, the compositions of the present invention may be used in conjunction with one or more of: a pharmaceutically acceptable carrier, a pharmaceutically acceptable diluent, a pharmaceutically acceptable excipient, a pharmaceutically acceptable adjuvant, a pharmaceutically active ingredient.

The pharmaceutical may be in the form of a liquid or as a solid - depending on the use and/or the mode of application and/or the mode of administration.

Medicaments

Compositions of the invention may take the form of medicaments.

The term "medicament" as used herein encompasses medicaments for both human and animal usage in human and veterinary medicine. In addition, the term "medicament" as used herein means any substance which provides a therapeutic, preventative and/or beneficial effect. The term "medicament" as used herein is not necessarily limited to substances which need marketing approval but may include substances which can be used in cosmetics, nutraceuticals, food (including feeds and beverages for example), probiotic cultures, and natural remedies. In addition, the term "medicament" as used herein encompasses a product designed for incorporation in animal feed, for example livestock feed and/or pet food.

Medical Foods

Compositions of the present invention may take the form of medical foods.

By "medical food" it is meant a food which is formulated to be consumed or administered with or without the supervision of a physician and which is intended for a specific dietary management or condition for which distinctive nutritional requirements, based on recognized scientific principles, are established by medical evaluation.

Dosage

The compositions of the present invention may comprise from 10⁶ to 10¹² colony forming units (CFU) of bacterial strain(s) per dose or per gram of composition, and more particularly from 10⁸ to 10¹² CFU of bacterial strain(s) per dose or per gram of composition. Optionally the compositions comprise about 10¹⁰ CFU of bacterial strain(s) per dose or per gram of composition. The bacterial strains(s) may be administered at a dosage from about 10⁶ to about 10¹² CFU of bacterial strain per dose, preferably about 10⁸ to about 10¹² CFU of bacterial strain per dose. By the term "per dose" it is meant that this number of bacteria is provided to a female mammal either per day or per intake, preferably per day. For example, if the bacteria are to be administered in a food product, for example in a yoghurt, then the yoghurt may contain from about 10⁶ to 10¹² CFU of the bacterial strain. Alternatively, however, this number of bacteria may be split into multiple administrations, each consisting of a smaller amount of microbial loading - so long as the overall amount of bacterial strain received by the female mammal in any specific time, for instance each 24 h period, is from about 10⁶ to about 10¹² CFU of bacteria, optionally 10⁸ to about 10¹² CFU of bacteria.

In accordance with the present invention an effective amount of at least one bacterial strain may be at least 10⁷ CFU of bacteria/dose, optionally from about 10⁸ to about 10¹² CFU of bacteria/dose, e.g., about 10¹⁰ CFU of bacteria/dose.

Effects/Female mammals/Medical indications

The term "female mammal", is used herein in its traditional meaning to refer to a mammal, including for example livestock (for example cattle, horses, pigs, and sheep) and humans having at least at birth the female reproductive system to produce egg cells or ova.

Prebiotics

In one embodiment, the bacterial strains and compositions of the present invention may further be combined or comprise one or more fibres and/or prebiotics.

Prebiotics are defined as a substrate that is selectively utilized by host microorganisms conferring a health benefit. These are generally ingredients that beneficially affect the health of the host by selectively stimulating the growth and/or activity of one or a limited number of bacteria, and thus improve host health. The prebiotic can be applied to oral route, but it can be also applied to other microbially colonized sites. Typically, prebiotics are carbohydrates (such as oligosaccharides), but the definition does not preclude non-carbohydrates, such as polyphenols, or polyunsaturated fatty acids or other ingredients that can be utilized selectively by a limited number of bacteria to confer a health benefit. The most prevalent forms of prebiotics are nutritionally classed as soluble fibres. To some extent, many forms of dietary fibres exhibit some level of prebiotic effect.

Examples of suitable prebiotics include alginate, xanthan, pectin, locust bean gum (LBG), inulin, guar gum, galacto-oligosaccharide (GOS), fructo-oligosaccharide (FOS), polydextrose 10 (i.e. Litesse®), lactitol, L-Arabinose, D-Xylose, L-Rhamnose, D-Mannose, L-Fucose, inositol, sorbitol, mannitol, xylitol, fructose, carrageenan, alginate, microcrystalline cellulose (MCC), betaine, lactosucrose, soybean oligosaccharides, isomaltulose (Palatinose TM), isomalto-oligosaccharides, gluco-oligosaccharides, xylooligosaccharides, manno-oligosaccharides, beta-glucans, cellobiose, raffinose, gentiobiose, melibiose, xylobiose, cyciodextrins, isomaltose, trehalose, stachyose, panose, pullulan, verbascose, galactomannans, (human) milk oligosaccharides and all forms of resistant starches.

Symptoms of menopause or climacteric symptoms

As used herein "symptom of menopause" refers to any symptom or condition specifically associated with the time when menstrual periods stop, such as permanently stop marking the end of reproduction in a female mammal, such as a female human, usually the time of the natural occurrence, but may also be induced prematurely, e.g. by surgery removing the ovaries or some types of chemotherapy also inducing early menopause either temporarily or permanently, or other conditions causing of a significant decrease in the ovaries' production of the hormones estrogen and progesterone.

The term is intended to include both peri- and postmenopausal symptoms. The perimenopause being the menopausal transition period leading up to complete menopause, i.e. a period with reduced synthesis of sex steroids. The "climacteric symptoms" are often referred to symptoms of perimenopausal women, typically in the months or years leading up to menopause.

Numbered embodiments of the invention:

1. A therapeutic composition comprising a combination of at least one bacterial strain of Bifidobacterium animalis subsp. lactis in combination with at least one bacterial strain selected from a strain of Lacticaseibacillus paracasei and/or a strain of Lactiplantibacillus plantarum for use in treating, preventing and/or alleviating one or more symptom of menopause or climacteric symptom in a female mammal, such as a female human.

2. The therapeutic composition for use according to embodiment 1, wherein the symptom of menopause or climacteric symptom is selected from the list consisting of vasomotor symptoms, including hot flushes, sweating, such as night sweat, sensations of heat, tachycardia, heart palpitations, chills, clamminess, headache, and changes in blood pressure; menopausal symptoms that may be associated with vasomotor symptoms or independent of vasomotor symptoms including disturbed sleep, insomnia, fatigue or tiredness, irritability, anxiousness, nervousness, poor memory, reduced concentration, dizziness, inattention, mood swings, feeling depressed or having low mood, lack of energy, perceived stress; weight gain and slowed metabolism, imbalanced blood glucose levels; genitourinary and sexual symptoms including vaginal dryness, vaginal discomfort, incontinence, frequent urination, dyspareunia, reduced libido; musculoskeletal symptoms such as myalgia, arthralgia, back pain, stiffness of joints, muscle and/or joint ache; gastrointestinal symptoms such as feeling bloated, constipation, gas, gastrointestinal pain, nausea, and other gastrointestinal discomfort. The therapeutic composition for use according to embodiment 2, wherein the symptom of menopause are vasomotor symptoms, such as hot flushes, sweating, such as night sweat, sensations of heat, tachycardia, heart palpitations, chills, clamminess, headache, and changes in blood pressure. The therapeutic composition for use according to embodiments 1-3, wherein the symptoms are associated with the perimenopause stage. The therapeutic composition for use according to embodiment 1-4, wherein the symptoms are associated with the post-menopause stage. The therapeutic composition for use according to embodiment 1-5, wherein the symptoms are associated with declining or fluctuating estrogen levels. The therapeutic composition for use according to embodiment 1-6, wherein the symptoms are associated with surgically induced menopause, such as hysterectomy and/or ovariectomy, and/or wherein the symptoms are induced by chemotherapy. The therapeutic composition for use according to embodiment 1-7, wherein the bacterial strain of Bifidobacterium animalis subsp. lactis is selected from Bifidobacterium animalis subsp. lactis deposited at DSMZ as DSM 15954 (BB-12® ), Bifidobacterium animalis subsp. lactis B420 deposited at DSMZ as DSM 32073, Bifidobacterium animalis subsp. lactis deposited at DSMZ as DSM 10140, Bifidobacterium animalis subsp. lactis HN019 (also known as DR.10™) deposited at DSMZ as DSM 17280), Bifidobacterium animalis subsp. lactis BI-04 (also known as DGCC2908 deposited at DSMZ as number DSM 33525), Bifidobacterium animalis subsp. lactis Bi-07 deposited at DSMZ as DSM 22878, Bifidobacterium animalis subsp lactis deposited as ATCC 27536, Bifidobacterium lactis NCC 2818 (CNCM 1-3446), and DN- 173-010 deposited at CNCM with accession number 1-2494. The therapeutic composition for use according to embodiment 1-8, wherein the bacterial strain of Lacticaseibacillus paracasei is selected from Lacticaseibacillus paracasei subsp. paracasei, L. casei 431®, Lacticaseibacillus paracasei ATCC 55544, Lacticaseibacillus paracasei JY062, Lacticaseibacillus paracasei PC804, Lacticaseibacillus paracasei F19®, Lacticaseibacillus paracasei subsp. paracasei LP-33®, Lacticaseibacillus paracasei ATCC 25302, Lacticaseibacillus paracasei strain GM-080, and the Lacticaseibacillus paracasei strain Lpc-37, registered at the DSMZ under deposit number DSM32661. The therapeutic composition for use according to embodiment 1-9, wherein the bacterial strain of Lactiplantibacillus plantarum is selected from Lactiplantibacillus plantarum GKM3, which is deposited in China General Microbiological Culture Collection Center (CGMCC) with a deposition number of CGMCC 14565, Lactiplantibacillus plantarum NK3 deposited at the Korean Microbial Conservation Center, under Accession No: KCCM12089P, Lactiplantibacillus plantarum strain ATCC202195, Lactiplantibacillus plantarum strain LP12418 deposited with the DSMZ under deposit number DSM 32655, Lactiplantibacillus plantarum strain LP12407 deposited with the DSMZ under deposit number DSM 32654, Lactiplantibacillus plantarum strain LP12151 deposited with the DSMZ under deposit number DSM 32721, Lactiplantibacillus plantarum strain deposited at the DSMZ under accession number DSM25833, Lactiplantibacillus plantarum Lp-115, deposited at DSMZ, under number DSM22266. The therapeutic composition for use according to embodiment 1-10, wherein the composition is in the form of a food product, a dietary supplement, functional food, or a pharmaceutically acceptable composition. The therapeutic composition for use according to embodiment 1-11, wherein the composition is a spray-dried or freeze-dried composition. The therapeutic composition for use according to embodiment 1-12, wherein the composition comprises a cryoprotectant. The therapeutic composition for use according to embodiment 1-13, wherein the bacterial strain of Bifidobacterium animalis subsp. lactis is present in the composition in an amount between 10⁶ and 10¹² CFU per dose. The therapeutic composition for use according to embodiment 1-14, wherein the bacterial strain of Lacticaseibacillus paracasei is present in the composition in an amount between 10⁶ and 10¹² CFU per dose. The therapeutic composition for use according to embodiment 1-15, wherein the bacterial strain of Lactiplantibacillus plantarum is present in the composition in an amount between 10⁶ and 10¹² CFU per dose. The therapeutic composition for use according to embodiment 1-16, which comprises or consists of at least one bacterial strain of Bifidobacterium animalis subsp. lactis in combination with at least one bacterial strain of Lacticaseibacillus paracasei. The therapeutic composition for use according to embodiment 1-16, which comprises or consists of at least one bacterial strain of Bifidobacterium animalis subsp. lactis in combination with at least one bacterial strain of Lactiplantibacillus plantarum. The therapeutic composition for use according to embodiment 1-16, which comprises or consists of at least one bacterial strain of Bifidobacterium animalis subsp. lactis in combination with at least one bacterial strain of Lacticaseibacillus paracasei or at least one bacterial strain of Lactiplantibacillus plantarum. A composition of a bacterial strain of Bifidobacterium animalis subsp. lactis in combination with a bacterial strain of Lacticaseibacillus paracasei and/or Lactiplantibacillus plantarum for use in reducing the need for estrogen replacement therapy, or for use in combination with estrogen replacement therapy, or other menopausal hormone replacement therapies. Use of a composition of a bacterial strain of Bifidobacterium animalis subsp. lactis in combination with a bacterial strain of Lacticaseibacillus paracasei and/or Lactiplantibacillus plantarum for the manufacture of a food product, a dietary supplement or a pharmaceutically acceptable formulation for use in treating, preventing and/or alleviating one or more symptom of menopause in a female mammal, such as a female human. Use of a composition of a bacterial strain of Bifidobacterium animalis subsp. lactis in combination with a bacterial strain of Lacticaseibacillus paracasei and/or Lactiplantibacillus plantarum for use in treating, preventing and/or alleviating one or more symptom of menopause in a female mammal, such as a female human. A method for treating, preventing and/or alleviating one or more symptom of menopause in a female mammal, such as a female human comprising the steps of: a) providing a composition of at least one bacterial strain of Bifidobacterium animalis subsp. lactis in combination with at least one bacterial strain selected from a strain of Lacticaseibacillus paracasei and/or a strain of Lactiplantibacillus plantarum,- and b) administering said composition of strains to said female mammal under conditions such that the risk of developing one or more symptom of menopause is reduced. The use according to embodiments 21 or 22 or method according to embodiment 23, wherein the composition of at least one bacterial strain of Bifidobacterium animalis subsp. lactis in combination with at least one bacterial strain selected from a strain of Lacticaseibacillus paracasei and/or a strain of Lactiplantibacillus plantarum is as defined in any one of embodiments 2-20. EXAMPLES

EXAMPLE 1

Animals and grouping

The study was conducted using three-month old female Sprague Dawley rats. They were divided into the following groups:

1. Sham-operated control group receiving vehicle (saline) p.o, n=10

2. OVX-group receiving vehicle (saline), p.o., n=10

3. OVX-group receiving reference compound estradiol (E2), s.c., n=10

4. OVX- group receiving B. lactis B420 + L. plantarum Lp-115, p.o., n=10

5. OVX- group receiving B. lactis BI-04 + L. plantarum Lp-115, p.o., n = 10

6. OVX- group receiving B. lactis B420 + L. paracasei Lpc-37, p.o., n = 10.

The bacterial strains used were deposited by DuPont Nutrition Biosciences ApS, in accordance with the Budapest Treaty at the Leibniz-Institut Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ), Inhoffenstrasse 7B, 38124 Braunschweig, Germany, where they are recorded under the following registration numbers:

B. lactis B420, also known as Bifidobacterium animalis subsp. lactis (strain) 420 (B420) commercially available from DuPont Nutrition Biosciences ApS, previously Danisco, and now International Flavors & Fragrances Inc. deposited on 30 June 2015 under registration number DSM 32073.

Lactiplantibacillus plantarum Lp-115 deposited on 9 February 2009 under registration number DSM 22266.

Bifidobacterium lactis BI-04 deposited on 19 May 2020 under registration number DSM 33525 Lacticaseibacillus paracasei Lpc-37 deposited on 5 October 2017 under registration number DSM 32661.

Prior to the start of the in-life phase, rats were allowed to acclimatize to the animal facility environment and were conditioned to the treatment over a period of two weeks. 2-4 rats were housed in each cage under controlled conditions of temperature (21°C ±3 °C) and light (12:12 h lig ht/dark cycle) and with tap water and diet ad libitum. The rats received standard rodent diet (2016/2916 Teklad Global Protein Rodent Diet) throughout the study.

Description of the ovariectomy procedure

Bilateral ovariectomy (OVX) was performed to study groups 2-6 on study day 0. Group 1 went through a sham operation. The animals were given preanesthetic analgesia (buprenorphine) at least 30-60 min prior the surgery. In addition, before the skin incision a s.c injection of local analgesia (Lidocaine 4-5 mg/kg) was provided. For the operation, the animals were anesthetized using isoflurane vaporizer and isoflurane inhalation. For the OVX procedure, small incisions in the skin and muscle layers on the dorsolateral site just below the ribs were made and the ovaries were pulled out on both sides. For sham operated rats after incisions to skin and muscle layer was made, the ovaries were visually located, inspected, and left intact. As a part of the recovery follow-up, animals received postsurgical analgesia for the three following days.

Interventions and dosing

Dosing intervention was started on day 1 after surgical operations and continued for 12 weeks (up to day 84). The vehicle was saline solution (0.9% NaCI), and it was administered by oral gavage to animals in groups 1 and 2. 170-estradiol (E2; cat #E1024; Sigma-Aldrich, St. Louis, MO, USA) was used as a reference compound and was administered to group 3. E2 solution was administered s.c. once a day in a dose of 4 pg/kg/day.

Different combinations of probiotics were given as oral gavage to groups 4, 5 and 6. The daily dose was lxlO⁹ CFU for each probiotic strain, total daily dose being 2xl0⁹ CFU.

• Group 4 received B. lactis B420 + L. plantarum Lp-115 (B420 + Lp-115)

• Group 5 received B. lactis BI-04 + L. plantarum Lp-115 (BI-04 + Lp-115)

• Group 6 received B. lactis B420 + L. paracasei Lpc-37 (B420 + Lpc-37)

Tail temperature measurement during the study

On study days -3 (baseline), 7, 14, 28, 56 and 84, tail skin temperature (T_taii) was measured. The measurement was performed in the evening at least one hour after beginning of the dark period, when lights in the animal room are switched off. Ttaii was measured using 153-IR.B Infrared Thermometer (Bioseb) designed for cutaneous temperature of small rodents. The IR. probe was applied about 1 cm from the base of the tail and at about 1 cm from the skin surface. This non-invasive measurement was performed 3 times per each individual rat and the average Ttaii was reported.

Evaluation of whole body and uterine weight

Animal weights (non-fasted) were followed once a week for 12 weeks of the study duration. At the end of in-life phase (at study day 85), body weight was collected, and animals were anesthetized for sacrifice. Following euthanasia, uterus was dissected and weighed to verify the impact of OVX.

Blood glucose measurement

Blood glucose levels were measured on study days at baseline (Day -3) and on days 14, 28, 56 and 84. Blood glucose concentrations were measured as fasted glucose levels in the morning using AlphaTrak 2 Meter Blood Glucose Monitoring System (Abbot Animal Health, Maidenhead, England). The glucose levels were measured from a drop of blood obtained from the tail vein. Animal was warmed before sampling to increase blood flow in the tail using a heating cabinet.

Statistical analysis

Statistical analyses were performed with the statistical software R. (version 4.2.2; 2022-10-31 ucrt, www.r-project.org).

The following comparisons between groups were made:

1. Sham + vehicle vs. OVX + vehicle

2. OVX + E2 vs. OVX + vehicle

3. OVX + probiotic groups vs. OVX vehicle group

4. Sham + vehicle vs. OVX + E2

Longitudinal data was analyzed using mixed models and model contrasts. Endpoint data was analyzed using ANOVA & Tukey's HSD post hoc test, or Kruskal-Wallis test & Dunn's test if the assumptions of ANOVA were not met.

Results

We show that probiotic combinations with bifidobacterial and lactobacilli strains were able to reduce the estrogen loss associated increase in tail skin temperature, a proxy measure for menopausal hot flushes. The reduction in T_taii was observed with all the tested probiotic combinations (B. animalis subsp. lactis B420 + L. plantarum Lp-115, B. animalis subsp. lactis BI-04 + L. plantarum Lp-115, and B. animalis subsp. lactis B420 + L. paracasei Lpc-37).

1 ) OVX Model verification

The OVX model was successfully established. The OVX rats receiving vehicle (OVX control) gained more weight than the sham operated animals as seen over the 12-week intervention period (Fig. 1, Table 1). Total body weight at sacrifice on day 85 of the OVX control group was also significantly higher as compared to the Sham group (Fig. 1). Treatment with estradiol (E2) significantly reduced the total body weight compared to the OVX animals receiving vehicle (Fig. 1, Table 1). The body weight of OVX animals treated with probiotic did not differ from OVX control group (Fig. 1, Table 1).

As a result of removal of the ovaries, the wet weight of the uterus of OVX animals was significantly reduced compared to the sham operated animals. Treatment with E2 counteracted the OVX induced changes in uterus weight (Fig. 2). Treatment with the probiotic combinations did not alter the uterus weight when compared to the OVX control group (Fig. 2).

2) Probiotics improve estrogen loss associated increase in tail skin temperature Tail skin temperature (Ttaii) was measured on days -3 (baseline), 7, 14, 28, 56 and 84. OVX animals receiving vehicle (OVX control) showed increased Ttaii after the first week of ovariectomy by about 2°C from the baseline and remained at that level throughout the study (Table 2). The OVX control group demonstrated significantly higher Ttaii compared to Sham group during the 12-week intervention (p=0.0029; Table 3). Treatment with E2 was able to counteract the OVX induced increase in Ttaii. Compared to OVX control group, E2 supplementation significantly reduced Ttaii (p=0.0045; Table 3).

OVX animals receiving any of the probiotic combinations showed a reduction in Ttaii towards the end of the study (Table 2). Compared to OVX control group, all three probiotic combinations showed reduction in Ttaii (B420 + Lp-115, p=0.023; BI-04 + Lp-115, p<0.001; B420 + Lpc-37, p=0.004; Table 3). After the 12-week intervention period, the reduction in Ttaii on day 84 was significant in OVX animals receiving B420 + Lpc-37 and BI-04 + Lp-115 (p<0.01 and p<0.05, respectively; Fig. 3) compared to the OVX control group receiving vehicle, and nearly significant in OVX animals receiving B420 + Lp-115 (p=0.0665, Fig. 3).

3) Probiotics reduce blood glucose in estrogen deficient subjects

Administration of probiotics to OVX animals resulted in reduction of blood glucose when compared to OVX control animals. After the 12-week supplementation on day 84, significant reduction in blood glucose was detected in OVX animals receiving B420 + Lp-115 (p=0.013; Table 4) and B420 + Lpc-37 (p=0.012; Table 4) when compared to animals in the OVX control group. The reduction in blood glucose was close to significant in BI-04 + Lp-115 supplemented animals vs. OVX control animals (p=0.144; Table 4).

1 ) OVX Model verification

Table 1. Body weight development and comparison³ between groups in body weight (g) during the 12-week intervention. Values represent the group average and standard deviation (SD). a The continuous data of the timepoints were modeled using a linear mixed-effects model and the comparisons were carried out using model contrasts, p-value when compared with OVX control group, comp=comparison group.

Notation: <= group smaller than the comparator group (comp), OVX group.

2 Probiotics attenuate ovariectomy induced increase in tail skin temperature.

Table 2. Tail temperature (°C) at the measured timepoints during the 12-week intervention.

Values represent the group average and standard deviation (SD).

***=p < 0.001; **=p < 0.01; *=p < 0.05; ^=0.0665, a statistical difference between the OVX group at the measured timepoints.

Table 3. Comparison³ between groups in tail skin temperature, T_taii, during the 12-week intervention. a The continuous data were modeled using a linear mixed-effects model and the comparisons were carried out using model contrasts. Notation: ***p < 0.001, **p < 0.01, *p < 0.05, ns=non-significant, '<’=former group smaller than latter.

3) Probiotics reduce blood glucose in estrogen deficient subjects

Table 4. Blood glucose on day 84 (group average and standard deviation, SD).

Statistical analysis was performed using ANOVA. As statistical differences were observed, the pairwise comparisons were performed using ANOVA contrasts. a p-value when compared with OVX control group, comp=comparison group.

*p < 0.05, ns=non-significant

Direction when compared to OVX control, <=smaller than comp

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Claims

1. A composition comprising a combination of at least one bacterial strain of Bifidobacterium an- imaiis subsp. lactis in combination with at least one bacterial strain selected from a strain of Lacticaseibacillus paracasei and/or a strain of Lactiplantibacillus plantarum for use in treating, preventing and/or alleviating one or more symptom of menopause or climacteric symptom in a female mammal, such as a female human.

2. The composition according to claim 1, wherein the symptom of menopause or climacteric symptom is selected from the list consisting of vasomotor symptoms, including hot flushes, sweating, such as night sweat, sensations of heat, tachycardia, heart palpitations, chills, clamminess, headache, and changes in blood pressure; menopausal symptoms that may be associated with vasomotor symptoms or independent of vasomotor symptoms including disturbed sleep, insomnia, fatigue or tiredness, irritability, anxiousness, nervousness, poor memory, reduced concentration, dizziness, inattention, mood swings, feeling depressed or having low mood, lack of energy, perceived stress; weight gain and slowed metabolism, imbalanced blood glucose levels; genitourinary and sexual symptoms including vaginal dryness, vaginal discomfort, incontinence, frequent urination, dyspareunia, reduced libido; musculoskeletal symptoms such as myalgia, arthralgia, back pain, stiffness of joints, muscle and/or joint ache; gastrointestinal symptoms such as feeling bloated, constipation, gas, gastrointestinal pain, nausea, and other gastrointestinal discomfort.

3. The composition according to claim 2, wherein the symptom of menopause are vasomotor symptoms, such as hot flushes, sweating, such as night sweat, sensations of heat, tachycardia, heart palpitations, chills, clamminess, headache, and changes in blood pressure.

4. The composition according to claims 1-3, wherein the symptoms are associated with the perimenopause stage, with the post-menopause stage, with declining or fluctuating estrogen levels, and/or with surgically induced menopause, such as hysterectomy and/or ovariectomy, and/or wherein the symptoms are induced by chemotherapy.

5. The composition according to claim 1-4, wherein the bacterial strain of Bifidobacterium ani- malis subsp. lactis is selected from Bifidobacterium animalis subsp. lactis deposited at DSMZ as DSM 15954 (BB-12® ), Bifidobacterium animalis subsp. lactis B420 deposited at DSMZ as DSM 32073, Bifidobacterium animalis subsp. lactis deposited at DSMZ as DSM 10140, Bifidobacterium animalis subsp. lactis HN019 (also known as DR.10™) deposited at DSMZ as DSM 17280), Bifidobacterium animalis subsp. lactis BI-04 (also known as DGCC2908 deposited at DSMZ as number DSM 33525), Bifidobacterium animalis subsp. lactis Bi-07 deposited at DSMZ as DSM 22878, Bifidobacterium animaiis subsp lactis deposited as ATCC 27536, Bifidobacterium lactis NCC 2818 (CNCM 1-3446), and DN-173-010 deposited at CNCM with accession number 1-2494.

6. The composition according to claim 1-5, wherein the bacterial strain of Lacticaseibacillus paracasei is selected from Lacticaseibacillus paracasei subsp. paracasei L. easel 431®, Lacticaseibacillus paracasei ATCC 55544, Lacticaseibacillus paracasei JY062, Lacticaseibacillus paracasei PC804, Lacticaseibacillus paracasei F19®, Lacticaseibacillus paracasei subsp. paracasei LP-33®, Lacticaseibacillus paracasei ATCC 25302, Lacticaseibacillus paracasei strain GM-080, and the Lacticaseibacillus paracasei strain Lpc-37, registered at the DSMZ under deposit number DSM32661.

7. The composition according to claim 1-6, wherein the bacterial strain of Lactiplantibacillus plantarum is selected from Lactiplantibacillus plantarum GKM3, which is deposited in China General Microbiological Culture Collection Center (CGMCC) with a deposition number of CGMCC 14565, Lactiplantibacillus plantarum NK3 deposited at the Korean Microbial Conservation Center, under Accession No: KCCM12089P, Lactiplantibacillus plantarum strain ATCC202195, Lactiplantibacillus plantarum strain LP12418 deposited with the DSMZ under deposit number DSM 32655, Lactiplantibacillus plantarum strain LP12407 deposited with the DSMZ under deposit number DSM 32654, Lactiplantibacillus plantarum strain LP12151 deposited with the DSMZ under deposit number DSM 32721, Lactiplantibacillus plantarum strain deposited at the DSMZ under accession number DSM25833, Lactiplantibacillusv plantarum Lp-115, deposited at DSMZ, under number DSM22266.

8. The composition according to claim 1-7, wherein the composition is in the form of a food product, a dietary supplement, functional food, or a pharmaceutically acceptable composition.

9. The composition according to claim 1-8, wherein the bacterial strain of Bifidobacterium animaiis subsp. lactis is present in the composition in an amount between 10⁶ and 10¹² CFU per dose, and/or the bacterial strain of Lacticaseibacillus paracasei is present in the composition in an amount between 10⁶ and 10¹² CFU per dose, and/or the bacterial strain of Lactiplantibacillus plantarum is present in the composition in an amount between 10⁶ and 10¹² CFU per dose.

10. The composition according to claim 1-9, which comprises or consists of at least one bacterial strain of Bifidobacterium animaiis subsp. lactis in combination with at least one bacterial strain of Lacticaseibacillus paracasei.

11. The composition according to claim 1-10, which comprises or consists of at least one bacterial strain of Bifidobacterium animaiis subsp. lactis in combination with at least one bacterial strain of Lactiplantibacillus plantarum.

12. The composition according to claim 1-11, which comprises or consists of at least one bacterial strain of Bifidobacterium animaiis subsp. lactis in combination with at least one bacterial strain of Lacticaseibacillus paracasei or at least one bacterial strain of Lactiplantibacillus plantarum.

13. A composition of a bacterial strain of Bifidobacterium animaiis subsp. lactis in combination with a bacterial strain of Lacticaseibacillus paracasei and/or Lactiplantibacillus plantarum for use in reducing the need for estrogen replacement therapy, or for use in combination with estrogen replacement therapy, or other menopausal hormone replacement therapies.

14. Use of a composition of a bacterial strain of Bifidobacterium animaiis subsp. lactis in combination with a bacterial strain of Lacticaseibacillus paracasei and/or Lactiplantibacillus plantarum for the manufacture of a food product, a dietary supplement or a pharmaceutically acceptable formulation for use in treating, preventing and/or alleviating one or more symptom of menopause in a female mammal, such as a female human.

15. The use according to claim 14, wherein the composition of at least one bacterial strain of Bifidobacterium animaiis subsp. lactis in combination with at least one bacterial strain selected from a strain of Lacticaseibacillus paracasei and/or a strain of Lactiplantibacillus plantarum is as defined in any one of claims 2-12.